Although much progress has been made deciphering the effects of anesthetics upon individual ion channels, identification of the neural substrates upon which anesthetics act to produce their behavioral effects remains as a challenge. Of the key components that characterize the anesthetized state, we focus on volatile anesthetic-induced hypnosis, defined as a lack of awareness to non-noxious stimuli. Understanding how anesthetics produce hypnosis has become more than a central question for neuroscience, as multiple reports over the past decade suggest that existing general anesthetics may annually harm a subset of the 40 million US patients who require anesthesia. One hypothetical alternative to anesthetic-induced unconsciousness is to generate a state of reversible physiological unconsciousness, such as sleep, in which the patient is locked out of access to the state of wakefulness. To determine whether existing volatile anesthetics cause their desirable hypnotic effects through interactions with endogenous sleep-promoting neural substrates, this proposal focuses on two emerging hypothalamic targets with proven ability to affect arousal state: the median preoptic nucleus, MnPO, and ventrolateral preoptic nucleus, VLPO. Depolarization of these two regions is respectively thought to underlie onset and maintenance of natural sleep. Our global hypothesis is that volatile anesthetics cause hypnosis by affecting VLPO and MnPO function. In Aim 1, we will show that exposure to hypnotic doses of volatile anesthetics activates VLPO and MnPO using c-Fos immunohistochemistry and slice electrophysiology. We will establish that exposure to a non-immobilizer fails to activate VLPO and MnPO in mice and in hypothalamic slices exposed ex vivo. In Aim 2, we will determine if the subset of neurons activated during natural sleep is the same subset activated by anesthetic exposure. In Aim 3, we will determine whether local destruction of VLPO and MnPO, microinjection of drugs that impair firing of VLPO and MnPO, or light-induced hyperpolarization of VLPO cause resistance to anesthetic hypnosis as determined by righting reflex assays and processed EEG entropy. Finally, we will determine if light-induced depolarization of VLPO causes hypersensitivity to anesthetic hypnosis. Cumulatively, these aims will determine whether or not volatile anesthetic-induced hypnosis is caused by recruitment of VLPO and/or MnPO. PUBLIC HEALTH RELEVANCE: Inhaled volatile anesthetics are safely used in the vast majority of the nation's 40 million patients who annually require general anesthesia; however, these drugs may harm a subset of vulnerable patients. Surprisingly, mechanisms of anesthetic-induced unconsciousness remain poorly understood. This research will evaluate the effects of volatile anesthetics upon hypothalamic sleep-promoting neurons to test the hypothesis that volatile anesthetic-induced unconsciousness is caused by recruitment of endogenous sleep- promoting substrates.